Substituted gamma lactams as therapeutic agents

ABSTRACT

Therapeutic compounds, compositions, medicaments, and methods are disclosed herein.

CROSS-REFERENCE TO RELATED APPLICATION

This application is based, and claims priority under 35 U.S.C. § 120 toU.S. Provisional Patent Application No. 60/777,506 filed on Feb. 28,2006, and which is incorporated herein by reference.

BRIEF DESCRIPTION OF THE DRAWING FIGURES

FIGS. 1-17 show examples of methods that can be used to prepare thecompounds disclosed herein.

DESCRIPTION OF THE INVENTION

Ocular hypotensive agents are useful in the treatment of a number ofvarious ocular hypertensive conditions, such as post-surgical andpost-laser trabeculectomy ocular hypertensive episodes, glaucoma, and aspresurgical adjuncts.

Glaucoma is a disease of the eye characterized by increased intraocularpressure. On the basis of its etiology, glaucoma has been classified asprimary or secondary. For example, primary glaucoma in adults(congenital glaucoma) may be either open-angle or acute or chronicangle-closure. Secondary glaucoma results from pre-existing oculardiseases such as uveitis, intraocular tumor or an enlarged cataract.

The underlying causes of primary glaucoma are not yet known. Theincreased intraocular tension is due to the obstruction of aqueous humoroutflow. In chronic open-angle glaucoma, the anterior chamber and itsanatomic structures appear normal, but drainage of the aqueous humor isimpeded. In acute or chronic angle-closure glaucoma, the anteriorchamber is shallow, the filtration angle is narrowed, and the iris mayobstruct the trabecular meshwork at the entrance of the canal ofSchlemm. Dilation of the pupil may push the root of the iris forwardagainst the angle, and may produce pupilary block and thus precipitatean acute attack. Eyes with narrow anterior chamber angles arepredisposed to acute angle-closure glaucoma attacks of various degreesof severity.

Secondary glaucoma is caused by any interference with the flow ofaqueous humor from the posterior chamber into the anterior chamber andsubsequently, into the canal of Schlemm. Inflammatory disease of theanterior segment may prevent aqueous escape by causing completeposterior synechia in iris bombe, and may plug the drainage channel withexudates. Other common causes are intraocular tumors, enlargedcataracts, central retinal vein occlusion, trauma to the eye, operativeprocedures and intraocular hemorrhage.

Considering all types together, glaucoma occurs in about 2% of allpersons over the age of 40 and may be asymptotic for years beforeprogressing to rapid loss of vision. In cases where surgery is notindicated, topical β-adrenoreceptor antagonists have traditionally beenthe drugs of choice for treating glaucoma.

Certain eicosanoids and their derivatives are currently commerciallyavailable for use in glaucoma management. Eicosanoids and derivativesinclude numerous biologically important compounds such as prostaglandinsand their derivatives. Prostaglandins can be described as derivatives ofprostanoic acid which have the following structural formula:

Various types of prostaglandins are known, depending on the structureand substituents carried on the alicyclic ring of the prostanoic acidskeleton. Further classification is based on the number of unsaturatedbonds in the side chain indicated by numerical subscripts after thegeneric type of prostaglandin [e.g. prostaglandin E₁ (PGE₁),prostaglandin E₂ (PGE₂)], and on the configuration of the substituentson the alicyclic ring indicated by α or β [e.g. prostaglandin F_(2α)(PGF_(2β))].

One embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

Another embodiment is a compound according to the formula

or a pharmaceutically acceptable salt or a prodrug thereof.

In another embodiment, said compound, salt thereof, and/or prodrugthereof is used to treat and/or prevent glaucoma and/or ocularhypertension in a mammal.

In another embodiment, said compound, and/or salt thereof, and/orprodrug thereof is used in the manufacture of a medicament for thetreatment and/or prevention of glaucoma and/or ocular hypertension in amammal.

Another embodiment is a composition comprising said compound, and/orsalt thereof, and/or prodrug thereof, wherein said composition isophthalmically acceptable.

Use of this compound in the treatment and/or prevention, and/or in themanufacture of a medicament for the treatment and/or prevention, of anydisease and/or condition mentioned herein as related to prostaglandinEP2 activity is also contemplated.

The compounds disclosed herein are useful for the prevention ortreatment of glaucoma or ocular hypertension in mammals, or for themanufacture of a medicament for the treatment of glaucoma or ocularhypertension. They are also useful for the treatment of those diseasesdisclosed in the art as being amenable to treatment by prostaglandin EP₂agonist, such as the ones listed previously.

A “pharmaceutically acceptable salt” is any salt that retains theactivity of the parent compound and does not impart any additionaldeleterious or untoward effects on the subject to which it isadministered and in the context in which it is administered compared tothe parent compound. A pharmaceutically acceptable salt also refers toany salt which may form in vivo as a result of administration of anacid, another salt, or a prodrug which is converted into an acid orsalt.

Pharmaceutically acceptable salts of acidic functional groups may bederived from organic or inorganic bases. The salt may comprise a mono orpolyvalent ion. Of particular interest are the inorganic ions, lithium,sodium, potassium, calcium, and magnesium. Organic salts may be madewith amines, particularly ammonium salts such as mono-, di- and trialkylamines or ethanol amines. Salts may also be formed with caffeine,tromethamine and similar molecules. Hydrochloric acid or some otherpharmaceutically acceptable acid may form a salt with a compound thatincludes a basic group, such as an amine or a pyridine ring.

A “prodrug” is a compound which is converted to a therapeutically activecompound after administration, and the term should be interpreted asbroadly herein as is generally understood in the art. While notintending to limit the scope of the invention, conversion may occur byhydrolysis of an ester group or some other biologically labile group.Generally, but not necessarily, a prodrug is inactive or less activethan the therapeutically active compound to which it is converted. Esterprodrugs of the compounds disclosed herein are specificallycontemplated. An ester may be derived from a carboxylic acid of C1 (i.e.the terminal carboxylic acid of a natural prostaglandin), or an estermay be derived from a carboxylic acid functional group on another partof the molecule, such as on a phenyl ring. While not intending to belimiting, an ester may be an alkyl ester, an aryl ester, or a heteroarylester. The term alkyl has the meaning generally understood by thoseskilled in the art and refers to linear, branched, or cyclic alkylmoieties. C₁₋₆ alkyl esters are particularly useful, where alkyl part ofthe ester has from 1 to 6 carbon atoms and includes, but is not limitedto, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl,t-butyl, pentyl isomers, hexyl isomers, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, and combinations thereof having from 1-6 carbonatoms, etc.

A metabolite is broadly defined as a compound which is formed in vivofrom the disclosed compound.

Those skilled in the art will readily understand that for administrationor the manufacture of medicaments the compounds disclosed herein can beadmixed with pharmaceutically acceptable excipients which per se arewell known in the art. Specifically, a drug to be administeredsystemically, it may be confected as a powder, pill, tablet or the like,or as a solution, emulsion, suspension, aerosol, syrup or elixirsuitable for oral or parenteral administration or inhalation.

For solid dosage forms or medicaments, non-toxic solid carriers include,but are not limited to, pharmaceutical grades of mannitol, lactose,starch, magnesium stearate, sodium saccharin, the polyalkylene glycols,talcum, cellulose, glucose, sucrose and magnesium carbonate. The soliddosage forms may be uncoated or they may be coated by known techniquesto delay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distcaratemay be employed. They may also be coated by the technique described inthe U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release. Liquid pharmaceuticallyadministrable dosage forms can, for example, comprise a solution orsuspension of one or more of the presently useful compounds and optionalpharmaceutical adjutants in a carrier, such as for example, water,saline, aqueous dextrose, glycerol, ethanol and the like, to therebyform a solution or suspension. If desired, the pharmaceuticalcomposition to be administered may also contain minor amounts ofnontoxic auxiliary substances such as wetting or emulsifying agents, pHbuffering agents and the like. Typical examples of such auxiliary agentsare sodium acetate, sorbitan monolaurate, triethanolamine, sodiumacetate, triethanolamine oleate, etc. Actual methods of preparing suchdosage forms are known, or will be apparent, to those skilled in thisart; for example, see Remington's Pharmaceutical Sciences, MackPublishing Company, Easton, Pa., 16th Edition, 1980. The composition ofthe formulation to be administered, in any event, contains a quantity ofone or more of the presently useful compounds in an amount effective toprovide the desired therapeutic effect.

Parenteral administration is generally characterized by injection,either subcutaneously, intramuscularly or intravenously. Injectables canbe prepared in conventional forms, either as liquid solutions orsuspensions, solid forms suitable for solution or suspension in liquidprior to injection, or as emulsions. Suitable excipients are, forexample, water, saline, dextrose, glycerol, ethanol and the like. Inaddition, if desired, the injectable pharmaceutical compositions to beadministered may also contain minor amounts of non-toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agentsand the like.

The amount of the presently useful compound or compounds administeredis, of course, dependent on the therapeutic effect or effects desired,on the specific mammal being treated, on the severity and nature of themammal's condition, on the manner of administration, on the potency andpharmacodynamics of the particular compound or compounds employed, andon the judgment of the prescribing physician. The therapeuticallyeffective dosage of the presently useful compound or compounds ispreferably in the range of about 0.5 or about 1 to about 100 mg/kg/day.

A liquid which is ophthalmically acceptable is formulated such that itcan be administered topically to the eye. The comfort should bemaximized as much as possible, although sometimes formulationconsiderations (e.g. drug stability) may necessitate less than optimalcomfort. In the case that comfort cannot be maximized, the liquid shouldbe formulated such that the liquid is tolerable to the patient fortopical ophthalmic use. Additionally, an ophthalmically acceptableliquid should either be packaged for single use, or contain apreservative to prevent contamination over multiple uses.

For ophthalmic application, solutions or medicaments are often preparedusing a physiological saline solution as a major vehicle. Ophthalmicsolutions should preferably be maintained at a comfortable pH with anappropriate buffer system. The formulations may also containconventional, pharmaceutically acceptable preservatives, stabilizers andsurfactants.

Preservatives that may be used in the pharmaceutical compositions of thepresent invention include, but are not limited to, benzalkoniumchloride, chlorobutanol, thimerosal, phenylmercuric acetate andphenylmercuric nitrate. A useful surfactant is, for example, Tween 80.Likewise, various useful vehicles may be used in the ophthalmicpreparations of the present invention. These vehicles include, but arenot limited to, polyvinyl alcohol, povidone, hydroxypropyl methylcellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl celluloseand purified water.

Tonicity adjustors may be added as needed or convenient. They include,but are not limited to, salts, particularly sodium chloride, potassiumchloride, mannitol and glycerin, or any other suitable ophthalmicallyacceptable tonicity adjustor.

Various buffers and means for adjusting pH may be used so long as theresulting preparation is ophthalmically acceptable. Accordingly, buffersinclude acetate buffers, citrate buffers, phosphate buffers and boratebuffers. Acids or bases may be used to adjust the pH of theseformulations as needed.

In a similar vein, an ophthalmically acceptable antioxidant for use inthe present invention includes, but is not limited to, sodiummetabisulfite, sodium thiosulfate, acetylcysteine, butylatedhydroxyanisole and butylated hydroxytoluene.

Other excipient components which may be included in the ophthalmicpreparations are chelating agents. A useful chelating agent is edetatedisodium, although other chelating agents may also be used in place orin conjunction with it.

The ingredients are usually used in the following amounts:

Ingredient Amount (% w/v) active ingredient about 0.001-5 preservative  0-0.10 vehicle   0-40 tonicity adjustor   1-10 buffer 0.01-10 pHadjustor q.s. pH 4.5-7.5 antioxidant as needed surfactant as neededpurified water as needed to make 100%

For topical use, creams, ointments, gels, solutions or suspensions,etc., containing the compound disclosed herein are employed. Topicalformulations may generally be comprised of a pharmaceutical carrier,cosolvent, emulsifier, penetration enhancer, preservative system, andemollient.

The actual dose of the active compounds of the present invention dependson the specific compound, and on the condition to be treated; theselection of the appropriate dose is well within the knowledge of theskilled artisan.

The compounds disclosed herein are also useful in combination with otherdrugs useful for the treatment of glaucoma or other conditions.

EXAMPLE 15-[(R)-1-(4-tert-Butyl-phenyl)-5-oxo-pyrrolidin-2-ylmethoxy]-pentanoicacid (4)

Step 1. Arylation of 1 to Give 2

A solution of amide 1 (3.30 g, 14.4 mmol) in 1,4-dioxane (25 mL) wasadded to a mixture of 4,5-bis(triphenylphosphino)-9,9-dimethylxanthene(xantphos, 600 mg, 1.04 mmol), Pd₂(dba)₃ (317 mg, 0.35 mmol) and Cs₂CO₃(6.46 g, 19.8 mmol). 1-Bromo-4-tert-butylbenzene (2.40 mL, 13.8 mmol)was added and the reaction mixture was purged with nitrogen. The mixturewas heated at reflux for 19 h, then cooled to rt. The reaction mixturewas then filtered through celite, washing with CH₂Cl₂, and the filtratewas concentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (10%→20% EtOAc/Hexane, gradient) afforded3.53 g (71%) of the desired product 2.

Step 2. Deprotection of 2 to Give 3

HF-pyridine (5 mL) was added to a solution of silyl ether 2 (3.53 g,9.76 mmol) in MeCN (20 mL) in a plastic bottle. The reaction was stirredat rt for 5 h, then was quenched with saturated aqueous NaHCO₃ (250 mL).The mixture was extracted with EtOAc (3×100 mL). The combined extractswere washed with brine (150 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo to yield 2.14 g (89%) of the desired product 3.

Step 3. Alkylation of 3 to Give the Ester of 4

Sodium hydride (11 mg, 0.46 mmol) was added to a solution of alcohol 3(100 mg, 0.40 mmol) in THF (3 mL) at 0° C. under nitrogen. After 1 h at0° C., methyl 5-bromovalerate (67 μL, 0.47 mmol) was added and thereaction was allowed to warm to rt. After 3 h, tlc analysis showedmostly starting alcohol remaining and another portion of bromide (67 μL,0.47 mmol) was added. After 22 h total reaction time, the reaction wasquenched with 1 N HCl and extracted with EtOAc (3×25 mL). Combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(40% EtOAc/Hexane→EtOAc, gradient) afforded 19 mg (13%) of the desiredester.

Step 4. Saponification to Give 4

Aqueous lithium hydroxide (1 N, 0.5 mL) was added to a solution of esterfrom step 3 above (12.3 mg, 0.034 mmol) in THF (0.7 mL). After 2.5 h atrt, the reaction was acidified with 0.25 M HCl (5 mL) then extractedwith CH₂Cl₂ (3×7 mL). Combined extracts were dried (Na₂SO₄), filteredand concentrated in vacuo to afford 10.2 mg (86%) of the title compound(4).

EXAMPLE 23-[(R)-1-(4-tert-Butyl-phenyl)-5-oxo-pyrrolidin-2-ylmethoxymethyl]-benzoicacid (5)

Step 1. Alkylation of 3 to Give the Ester of 5

Potassium hydride (23.4 mg, 0.58 mmol) and 18-crown-6 (167 mg, 0.63mmol) were added sequentially to a solution of alcohol 3 (130 mg, 0.53mmol) in THF (3 mL) at 0° C. After 1 h at 0° C., a solution of methyl3-(chloromethyl)benzoate (prepared from the corresponding acid chloride,pyridine and methanol: see J. Org. Chem. 1988, 53, 2548-2552; 116 mg,0.63 mmol) in THF (1.5 mL) was added via cannula and the reaction wasallowed to warm to rt. After 22.5 h, the reaction was quenched with 0.1N HCl (10 mL) and extracted with EtOAc (3×15 mL). Combined extracts werewashed with saturated aqueous NaHCO₃ (15 mL) and brine (15 mL) thendried (Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by flash column chromatography on silica gel (30%→50%EtOAc/Hexane, gradient) afforded 66 mg (32%) of the desired ester.

Step 2. Saponification to Give 5

Aqueous lithium hydroxide (1 N, 0.4 mL) was added to a solution of esterfrom step 1 above (33.5 mg, 0.085 mmol) in THF (0.75 mL). After 3.5 h atrt, the reaction was acidified with 0.25 M HCl (5 mL) then extractedwith CH₂Cl₂ (3×10 mL). Combined extracts were dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (2% MeOH/CH₂Cl₂), followed by preparativethin layer chromatography (10% MeOH/CH₂Cl₂) afforded 6.6 mg (20%) of thetitle compound (5).

EXAMPLE 35-[(R1)-1-(4-tert-Butyl-phenyl)-5-oxo-pyrrolidin-2-ylmethoxymethyl]-furan-2-carboxylicacid (6)

Step 1. Alkylation of 3 to Give the Ester of 6

Potassium hydride (27 mg, 0.67 mmol) and 18-crown-6 (193 mg, 0.73 mmol)were added sequentially to a solution of alcohol 3 (150 mg, 0.61 mmol)in THF (4 mL) at 0° C. After 1 h at 0° C., a solution of ethyl5-chloromethylfuran-2-carboxylate (commercially available from AldrichChemical Company, 138 mg, 0.73 mmol) in THF (1 mL) was added via cannulaand the reaction was allowed to warm to rt. After 18.5 h, the reactionwas quenched with 0.25 N HCl (10 mL) and extracted with EtOAc (3×15 mL).Combined extracts were washed with brine (20 mL) then dried (Na₂SO₄),filtered and concentrated in vacuo. Purification of the residue by flashcolumn chromatography on silica gel (20%→50% EtOAc/Hexane, gradient)afforded 78 mg (32%) of the desired ester.

Step 2. Saponification to Give 6

Aqueous lithium hydroxide (1 N, 0.5 mL) was added to a solution of esterfrom step 1 above (66.7 mg, 0.17 mmol) in THF (0.5 mL). After 3 h at rt,the reaction was acidified with 1 N HCl (2 mL) then extracted withCH₂Cl₂ (3×10 mL). Combined extracts were dried (Na₂SO₄), filtered andconcentrated in vacuo to afford 54.4 mg (88%) of the title compound (6).

EXAMPLE 45-[(R)-1-(4-tert-Butyl-phenyl)-5-oxo-pyrrolidin-2-ylmethoxymethyl]-thiophene-2-carboxylicacid (7)

Step 1. Alkylation of 3 to Give the Ester of 7

Potassium hydride (25.2 mg, 0.63 mmol) and 18-crown-6 (181 mg, 0.68mmol) were added sequentially to a solution of alcohol 3 (140 mg, 0.57mmol) in THF (4 mL) at 0° C. After 1.5 h at 0° C., a solution of methyl5-chloromethylthiophene-2-carboxylate (prepared according to theprocedures described in WO2004/037808; 130 mg, 0.68 mmol) in THF (1.5mL) was added via cannula and the reaction was allowed to warm to rt.After 20 h, the reaction was quenched with 0.25 N HCl (15 mL) andextracted with EtOAc (3×20 mL). Combined extracts were washed with brine(30 mL) then dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(20%→50% EtOAc/Hexane, gradient) afforded 40.7 mg (18%) of the desiredester.

Step 2. Saponification to Give 7

Aqueous lithium hydroxide (1 N, 0.4 mL) was added to a solution of esterfrom step 1 above (37 mg, 0.092 mmol) in THF (0.75 mL). After 18 h atrt, the reaction was acidified with 1 N HCl (7 mL) then extracted withCH₂Cl₂ (3×10 mL). Combined extracts were dried (Na₂SO₄), filtered andconcentrated in vacuo to afford 22.3 mg (62%) of the title compound (7).

EXAMPLE 57-[(S)-1-(4-tert-Butyl-phenyl)-5-oxo-pyrrolidin-2-yl]-heptanoic acid(10)

Step 1. Oxidation of 3 to Give Aldehyde 8

Molecular sieves (4 Å, 300 mg), 4-methylmorpholine N-oxide (427 mg, 3.64mmol) and tetrapropylammonium perruthenate (250 mg, 0.71 mmol) wereadded sequentially to a solution of alcohol 3 (600 mg, 2.43 mmol) inCH₂Cl₂ (15 mL) at rt. After 23 h, the reaction mixture was filteredthrough celite, washing with CH₂Cl₂ (10 mL). The filtrate wasconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (CH₂Cl₂→10% EtOAc/CH₂Cl₂, gradient)afforded 92 mg (15%) of the desired aldehyde 8.

Step 2. Wittig Reaction of 8 to Give 9

Potassium bis(trimethylsilyl)amide (0.5 M in PhMe, 1.92 mL, 0.96 mmol)was added to a solution of aldehyde 8 (86 mg, 0.35 mmol) in THF (2 mL)at rt. After 15 min at rt, the reaction mixture was cooled to −55° C.for 10 min before a solution of 5-carboxypentyltriphenylphosphoniumbromide (207 mg, 0.45 mmol) was added via cannula. After 10 min at −55°C., the reaction was allowed to warm to rt. After 18 h at rt, thereaction was quenched with saturated aqueous NH₄Cl (15 mL) and extractedwith EtOAc (3×15 mL). Combined extracts were washed with brine (20 mL),dried (Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by preparative thin layer chromatography (5% MeOH/CH₂Cl₂)afforded 10.5 mg (9%) of desired alkene 9.

Step 3. Hydrogenation of 9 to Give 10

Palladium on carbon (10 wt. %, 2 mg) was added to a solution of alkene 9(5.8 mg, 0.017 mmol) in MeOH (1 mL). A hydrogen atmosphere wasestablished by evacuating and refilling with hydrogen (3×) and thereaction mixture was stirred under a balloon of hydrogen for 18 h. Thereaction mixture was filtered through celite, washing with MeOH, and thefiltrate was concentrated in vacuo to afford 4.1 mg (70%) of the titlecompound (10).

EXAMPLE 65-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-furan-2-carboxylicacid (17)

Step 1. Arylation of 1 to Give 12

A solution of amide 1 (2.89 g, 12.60 mmol) in 1,4-dioxane (20 mL)followed by a solution of 1-(4-methoxybenzyloxymethyl)-4-bromobenzene(11: for synthesis, see Allergan docket #17693; 3.88 g, 12.63 mmol) wereadded sequentially to a mixture of xantphos (877 mg, 1.52 mmol),Pd₂(dba)₃ (463 mg, 0.51 mmol) and Cs₂CO₃ (3.2 g, 9.82 mmol) via cannula.The reaction mixture was purged with nitrogen and then heated at refluxfor 22 h. The reaction mixture was allowed to cool to rt then filteredthrough celite, washing with CH₂Cl₂, and the filtrate was concentratedin vacuo. Purification of the residue by flash column chromatography onsilica gel (5%→25% EtOAc/Hexane, gradient) afforded 1.70 g (30%) ofdesired product 12.

Step 2. Deprotection of 12 to Give 13

HF-pyridine (5 mL) was added to a solution of silyl ether 12 (1.38 g,3.03 mmol) in MeCN (15 mL) in a plastic bottle at 0° C. The reaction wasstirred at 0° C. for 3 h, then was quenched with saturated aqueousNaHCO₃ (250 mL). The mixture was extracted with EtOAc (3×100 mL). Thecombined extracts were washed with brine (100 mL) then dried (Na₂SO₄),filtered and concentrated in vacuo. Purification of the residue by flashcolumn chromatography on silica gel (1%→3% MeOH/CH₂Cl₂, gradient)afforded 464 mg (45%) of desired alcohol 13.

Step 3. Alkylation of Alcohol 13 to Give 14

Potassium hydride (44 mg, 1.10 mmol) and 18-crown-6 (365 mg, 1.38 mmol)were added sequentially to a solution of alcohol 13 (315 mg, 0.92 mmol)in THF (4 mL) at 0° C. After 1 h at 0° C., ethyl5-chloromethylfuran-2-carboxylate (0.28 mL, 1.82 mmol) was added and thereaction was allowed to warm to rt. After 22 h, the reaction wasquenched with 0.5 N HCl (20 mL) and extracted with EtOAc (3×25 mL).Combined extracts were washed with brine (50 mL) then dried (Na₂SO₄),filtered and concentrated in vacuo. Purification of the residue by flashcolumn chromatography on silica gel (20% EtOAc/Hexane→EtOAc, gradient)afforded 148 mg (32%) of desired product 14.

Step 4. Oxidative Deprotection of 14 to Give 15 and 16

2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ, 82 mg, 0.36 mmol) wasadded to a mixture of 14 (143 mg, 0.29 mmol) in CH₂Cl₂ (4 mL) and water(0.2 mL). After 3 h, tlc indicated that starting material remained andanother portion of DDQ (82 mg, 0.36 mmol) was added. After a further1.25 h, the reaction was quenched with saturated aqueous NaHCO₃ (20 mL).The mixture was extracted with EtOAc (3×20 mL). The combined extractswere washed with brine (20 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (CH₂Cl₂→3% MeOH/CH₂Cl₂, gradient) afforded38 mg (35%) of the desired alcohol 15 and 61 mg of impure aldehyde 16.Aldehyde 16 was further purified by preparative thin layerchromatography (5% MeOH/CH₂Cl₂) to afford 48.7 mg (45%) of aldehyde 16.

Step 5. Oxidation of 15 to Give 16

Molecular sieves (4 Å, 3 mg), 4-methylmorpholine N-oxide (12.6 mg, 0.11mmol) and tetrapropylammonium perruthenate (2.5 mg, 0.007 mmol) wereadded sequentially to a solution of alcohol 15 (26.8 mg, 0.072 mmol) inCH₂Cl₂ (1.5 mL) at rt. After 20 min, the reaction mixture was filteredthrough celite, washing with CH₂Cl₂ (5 mL). The filtrate wasconcentrated in vacuo. Purification of the residue by preparative thinlayer chromatography (5% MeOH/CH₂Cl₂) afforded 9.6 mg (36%) of thedesired aldehyde 16.

Step 6. Grignard Reaction with 16 to Give the Ester of 17

Pentyl magnesium bromide (2.0 M in Et₂O, 32 μL, 0.064 mmol) was added toa solution of aldehyde 16 (21.7 mg, 0.058 mmol) in THF (0.4 mL) at −40°C. under nitrogen. After 25 min, the reaction was quenched withsaturated aqueous NH₄Cl and extracted with CH₂Cl₂ (3×7 mL). Combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by preparative thin layer chromatography (5%MeOH/CH₂Cl₂) afforded 10.6 mg (41%) of the desired ester.

Step 7. Saponification to Give 17

Aqueous lithium hydroxide (1 N, 0.1 mL) was added to a solution of esterfrom step 6 above (8.8 mg, 0.02 mmol) in THF (0.2 mL). After 1 h at rt,the reaction was acidified with 0.5 N HCl (1 mL) then extracted withCH₂Cl₂ (3×7 mL). Combined extracts were dried (Na₂SO₄), filtered andconcentrated in vacuo to afford 8.2 mg (99%) of the title compound (17).

EXAMPLE 75-{(R)-1-[4-(1-Hydroxy-2-methyl-propyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-furan-2-carboxylicacid (18)

Step 1. Grignard Reaction with 16 to Give the Ester of 18

Isopropyl magnesium chloride (2.0 M in THF, 31 μL, 0.062 mmol) was addedto a solution of aldehyde 16 (20.5 mg, 0.055 mmol) in THF (0.4 mL) at−40° C. under nitrogen. After 35 min, the reaction was quenched withsaturated aqueous NH₄Cl and extracted with CH₂Cl₂ (3×7 mL). Combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by preparative thin layer chromatography (5%MeOH/CH₂Cl₂) afforded 5 mg (22%) of the desired ester.

Step 2. Saponification to Give 18

Aqueous lithium hydroxide (1 N, 0.05 mL) was added to a solution of theester from step 1 above (3.1 mg, 0.007 mmol) in THF (0.15 mL). After 1 hat rt, the reaction was acidified with 0.2 N HCl (1 mL) then extractedwith CH₂Cl₂ (3×7 mL). Combined extracts were dried (Na₂SO₄), filteredand concentrated in vacuo to afford 2.5 mg (86%) of the title compound(18).

EXAMPLE 85-{(R)-1-[4-(1-Hydroxy-2-phenyl-ethyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-furan-2-carboxylicacid (19)

Step 1. Grignard Reaction with 16 to Give the Ester of 19

Benzyl magnesium chloride (2.0 M in THF, 14 μL, 0.028 mmol) was added toa solution of aldehyde 16 (9.6 mg, 0.026 mmol) in THF (0.3 mL) at −40°C. under nitrogen. After 45 min, the reaction was warmed to 0° C. After25 min at 0° C., the reaction was quenched with saturated aqueous NH₄Cland extracted with CH₂Cl₂ (3×7 mL). Combined extracts were dried(Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by preparative thin layer chromatography (7% MeOH/CH₂Cl₂)afforded 3.3 mg (28%) of the desired ester.

Step 2. Saponification to Give 19

Aqueous lithium hydroxide (1 N, 0.05 mL) was added to a solution of theester from step 1 above (2.4 mg, 0.005 mmol) in THF (0.15 mL). After 2.5h at rt, the reaction was acidified with 0.2 N HCl (1 mL) then extractedwith CH₂Cl₂ (3×7 mL). Combined extracts were dried (Na₂SO₄), filteredand concentrated in vacuo to afford 2.2 mg (98%) of the title compound(19)

EXAMPLE 95-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (23)

Step 1. Alkylation of 13 to Give 20

Potassium hydride (55.5 mg, 1.38 mmol) and 18-crown-6 (456 mg, 1.73mmol) were added sequentially to a solution of alcohol 13 (394 mg, 1.15mmol) in THF (5 mL) at 0° C. After 1 h at 0° C., a solution of methyl5-chloromethylthiophene-2-carboxylate (439 mg, 2.30 mmol) in THF (2 mL)was added via cannula and the reaction was allowed to warm to rt. After19 h, tlc analysis showed starting material remained. Another portion ofKH (20 mg, 0.50 mmol) was added and the reaction was heated at 50° C.After 2 h at 50° C., the reaction was cooled and quenched with 0.5 N HCl(20 mL) and extracted with EtOAc (3×25 mL). Combined extracts werewashed with brine (50 mL) then dried (Na₂SO₄), filtered and concentratedin vacuo. Purification of the residue by flash column chromatography onsilica gel (15% EtOAc/Hexane→EtOAc, gradient) afforded 108 mg (19%) ofdesired product 20.

Step 2. Oxidative Deprotection of 20 to Give 21 and 22

DDQ (91 mg, 0.40 mmol) was added to a mixture of 20 (98 mg, 0.20 mmol)in CH₂Cl₂ (3 mL) and water (0.15 mL). After 4.5 h, the reaction wasquenched with saturated aqueous NaHCO₃ (15 mL) and extracted with EtOAc(3×25 mL). Combined extracts were washed with brine (40 mL) then dried(Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by preparative thin layer chromatography (5% MeOH/CH₂Cl₂)afforded 14.4 mg (19%) of alcohol 21 and 16.2 mg (22%) of aldehyde 22.

Step 3. Grignard Reaction with 22 to Give the Ester of 23

Pentyl magnesium bromide (2.0 M in Et₂O, 22 μL, 0.044 mmol) was added toa solution of aldehyde 22 (11 mg, 0.029 mmol) in THF (0.2 mL) at −40° C.under nitrogen. After 1.5 h, the reaction was quenched with saturatedaqueous NH₄Cl and extracted with CH₂Cl₂ (3×7 mL). Combined extracts weredried (Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by preparative thin layer chromatography (5% MeOH/CH₂Cl₂)afforded 4.8 mg (37%) of the desired ester.

Step 4. Saponification to Give 23

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 3 above (3.6 mg, 0.008 mmol) in MeCN (0.1 mL) and pH7.2 buffer (2.5 mL). After 16.5 h at rt, the reaction was diluted withMeCN (7 mL) and concentrated in vacuo. The residue was suspended inCH₂Cl₂ and filtered through a cotton plug. The filtrate was concentratedin vacuo to afford 2.0 mg (57%) of the title compound (23).

EXAMPLE 105-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (28)

Step 1. Arylation of 1 to Give 24

A solution of amide 1 (3.37 g, 14.7 mmol) in 1,4-dioxane (30 mL) wasadded to a mixture of Pd₂(dba)₃ (540 mg, 0.59 mmol), xantphos (1.02 g,1.76 mmol) and Cs₂CO₃ (5.74 g, 17.6 mmol). A solution of1-(1-(4-methoxybenzyloxyhexyl)-4-bromobenzene (preparation 1, 4.99 g,13.22 mmol) in 1,4-dioxane (30 mL) was added via cannula, followed by anadditional 40 mL of 1,4-dioxane. The reaction mixture was purged withnitrogen then heated at reflux overnight. After 20 h, the reaction wascooled to rt and filtered through celite, washing with CH₂Cl₂. Thefiltrate was concentrated in vacuo and the residue was purified by flashcolumn chromatography on silica gel (5%→30% EtOAc/Hexane, gradient) toafford 5.79 g (83%) of the desired product 24.

Step 2. Deprotection of 24 to Give 25

HF-pyridine (7 mL) was added to a solution of silyl ether 24 (4.05 g,7.72 mmol) in MeCN (40 mL) in a plastic bottle at 0° C. The reaction wasstirred at 0° C. for 1 h, then was quenched with saturated aqueousNaHCO₃ (300 mL). The mixture was extracted with EtOAc (3×150 mL). Thecombined extracts were washed with brine (200 mL) then dried (Na₂SO₄),filtered and concentrated in vacuo. Purification of the residue by flashcolumn chromatography on silica (CH₂Cl₂→3% MeOH/CH₂Cl₂, gradient)afforded 2.3 g (72%) of the desired alcohol 25.

Step 3. Alkylation of 25 to Give 26

Potassium hydride (155 mg, 3.86 mmol) was added to a solution of alcohol25 (1.22 g, 2.97 mmol) in THF (7 mL) at 0° C. After 15 min at 0° C.,18-crown-6 (1.02 g, 3.86 mmol) was added. After 45 min longer at 0° C.,a solution of isopropyl 5-chloromethylthiophene-2-carboxylate(preparation 2, 650 mg, 2.97 mmol) in THF (5 mL) was added via cannula.Potassium iodide (50 mg, 0.30 mmol) was added and the reaction wasallowed to warm to rt. After 20 h, the reaction was quenched with 0.5 NHCl (70 mL) and extracted with EtOAc (3×100 mL). Combined extracts werewashed with brine (100 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (20% EtOAc/Hexane→EtOAc, gradient) afforded296 mg (17%) of desired product 26 along with 747 mg (61%) of recoveredstarting alcohol 25.

Step 4. Oxidative Deprotection of 26 to Give 27 and 28

DDQ (93 mg, 0.41 mmol) was added to a solution of 26 (220 mg, 0.37 mmol)in CH₂Cl₂ (4 mL) and water (0.2 mL) at 0° C. under nitrogen. After 35min, the reaction was quenched with saturated aqueous NaHCO₃ (30 mL) andextracted with EtOAc (3×30 mL). Combined extracts were washed with brine(50 mL) then dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(20%→70% EtOAc/Hexane, gradient) afforded 13 mg (7%) of ketone 27 and108 mg (62%) of the title compound (28).

EXAMPLE 113-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-benzoicacid (31)

Step 1. Alkylation of 13 to Give 29

Potassium hydride (16 mg, 0.39 mmol) was added to a solution of alcohol13 (112 mg, 0.33 mmol) in THF (1.0 mL) at 0° C. After 1 h at 0° C.,18-crown-6 (114 mg, 0.43 mmol), potassium iodide (5 mg, 0.03 mmol) and asolution of methyl 3-chloromethylbenzoate (121 mg, 0.66 mmol) in THF(0.5 mL) were added sequentially. The reaction was allowed to warm tort. After 19 h, the reaction was quenched with 0.1 N HCl (10 mL) andextracted with EtOAc (3×10 mL). Combined extracts were washed with brine(15 mL) then dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(20% EtOAc/Hexane→EtOAc, gradient) afforded 23 mg (14%) of desiredproduct 29.

Step 2. Oxidative Deprotection of 29 to Give 30

DDQ (23 mg, 0.10 mmol) was added to a mixture of 29 (23 mg, 0.047 mmol)in CH₂Cl₂ and water (20:1, 0.25 mL). After 3.75 h, the reaction wasquenched with saturated aqueous NaHCO₃ (10 mL) and extracted with EtOAc(3×7 mL). Combined extracts were washed with brine (10 mL) then dried(Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by preparative thin layer chromatography (80% EtOAc/Hex)afforded 13 mg (58%) of aldehyde 30.

Step 3. Grignard Reaction with 30 to Give the Ester of 31

Pentyl magnesium bromide (2.0 M in Et₂O, 50 μL, 0.10 mmol) was added toa solution of aldehyde 30 (12.4 mg, 0.034 mmol) in THF (0.1 mL) at −40°C. under nitrogen. After 1 h, the reaction was quenched with saturatedaqueous NH₄Cl (7 mL) and extracted with CH₂Cl₂ (3×7 mL). Combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by preparative thin layer chromatography (5%MeOH/CH₂Cl₂) afforded 8.6 mg (58%) of the desired ester.

Step 4. Saponification to Give 31

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 3 above (7.4 mg, 0.017 mmol) in MeCN (0.1 mL) and pH7.2 buffer (2.5 mL). After 18 h at rt, the reaction was diluted withMeCN (7 mL) and concentrated in vacuo. Purification of the residue bypreparative thin layer chromatography (5% MeOH/CH₂Cl₂) afforded 1.5 mg(21%) of the title compound (31).

EXAMPLE 125-{(R)-1-[4-(1-Hydroxy-pentyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (32)

Step 1. Grignard Reaction with 22 to Give the Ester of 32

n-Butyl magnesium chloride (2.0 M in THF, 41 μL, 0.082 mmol) was addedto a solution of aldehyde 22 (20.2 mg, 0.054 mmol) in THF (0.1 mL) at−40° C. under nitrogen. After 1 h, the reaction was quenched withsaturated aqueous NH₄Cl (10 mL) and extracted with CH₂Cl₂ (3×7 mL).Combined extracts were dried (Na₂SO₄), filtered and concentrated invacuo. Purification of the residue by preparative thin layerchromatography (5% MeOH/CH₂Cl₂) afforded 12.3 mg (53%) of the desiredester.

Step 2. Saponification to Give 32

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 1 above (11.2 mg, 0.026 mmol) in MeCN (0.1 mL) andpH 7.2 buffer (3.0 mL). After 19 h at rt, the reaction was diluted withMeCN (10 mL) and concentrated in vacuo. The residue was suspended in 5%MeOH/CH₂Cl₂ and filtered through a cotton plug. The filtrate wasconcentrated in vacuo to afford 10.7 mg (99%) of the title compound(32).

EXAMPLE 135-{(R)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (33)

Step 1. Grignard reaction with 22 to give the ester of 33

n-Hexyl magnesium bromide (2.0 M in Et₂O, 100 μL, 0.20 mmol) was addedto a solution of aldehyde 22 (24.6 mg, 0.054 mmol) in THF (0.12 mL) at−40° C. under nitrogen. After 1.5 h, the reaction was quenched withsaturated aqueous NH₄Cl (10 mL) and extracted with CH₂Cl₂ (3×7 mL).Combined extracts were dried (Na₂SO₄), filtered and concentrated invacuo. Purification of the residue by preparative thin layerchromatography (5% MeOH/CH₂Cl₂) afforded 16.3 mg (54%) of the desiredester.

Step 2. Saponification to Give 33

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 1 above (13 mg, 0.028 mmol) in MeCN (0.1 mL) and pH7.2 buffer (3.0 mL). After 18 h at rt, the reaction was diluted withMeCN (10 mL) and concentrated in vacuo. The residue was suspended in 5%MeOH/CH₂Cl₂ and filtered through a cotton plug. The filtrate wasconcentrated in vacuo to afford 11 mg (87%) of the title compound (33).

EXAMPLE 145-{(R)-1-[4-(1-Hydroxy-butyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (34)

Step 1. Grignard Reaction with 22 to Give the Ester of 34

n-Propyl magnesium chloride (2.0 M in Et₂O, 92 μL, 0.18 mmol) was addedto a solution of aldehyde 22 (22.9 mg, 0.061 mmol) in THF (0.12 mL) at−40° C. under nitrogen. After 1.75 h, the reaction was quenched withsaturated aqueous NH₄Cl (10 mL) and extracted with CH₂Cl₂ (3×7 mL).Combined extracts were dried (Na₂SO₄), filtered and concentrated invacuo. Purification of the residue by preparative thin layerchromatography (5% MeOH/CH₂Cl₂) afforded 13 mg (51%) of the desiredester.

Step 2. Saponification to Give 34

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 1 above (10.8 mg, 0.026 mmol) in MeCN (0.1 mL) andpH 7.2 buffer (3.0 mL). After 17 h at rt, the reaction was diluted withMeCN (10 mL) and concentrated in vacuo. The residue was suspended in 5%MeOH/CH₂Cl₂ and filtered through a cotton plug. The filtrate wasconcentrated in vacuo to afford 10.4 mg (99%) of the title compound(34).

EXAMPLE 155-{(R)-1-[4-(1-Hydroxy-propyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (35)

Step 1. Grignard Reaction with 22 to Give the Ester of 35

Ethyl magnesium chloride (2.0 M in Et₂O, 24 μL, 0.048 mmol) was added toa solution of aldehyde 22 (5.8 mg, 0.016 mmol) in THF (0.1 mL) at −40°C. under nitrogen. After 1.25 h, the reaction was quenched withsaturated aqueous NH₄Cl (5 mL) and extracted with CH₂Cl₂ (3×5 mL).Combined extracts were dried (Na₂SO₄), filtered and concentrated invacuo. Purification of the residue by preparative thin layerchromatography (5% MeOH/CH₂Cl₂) afforded 2.5 mg (40%) of the desiredester.

Step 2. Saponification to Give 35

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofthe ester from step 1 above (2.8 mg, 0.007 mmol) in MeCN (0.1 mL) and pH7.2 buffer (2.5 mL). After 17 h at rt, the reaction was diluted withMeCN (10 mL) and concentrated in vacuo. The residue was suspended in 5%MeOH/CH₂Cl₂ and filtered through a cotton plug. The filtrate wasconcentrated in vacuo to afford 2.7 mg (99%) of the title compound (35).

EXAMPLE 16 5-((E andZ)-3-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-allyl)-thiophene-2-carboxylicacid (41)

Step 1. Oxidation of 25 to Give Aldehyde 36

Dess-Martin periodinane (1.63 g, 3.83 mmol) was added to a solution ofalcohol 25 (1.43 g, 3.48 mmol) in CH₂Cl₂ (12 mL) at rt under nitrogen.After 1 h at rt the reaction was quenched with saturated aqueous NaHCO₃and saturated aqueous NaHSO₃ (1:1100 mL). The mixture was extracted withCH₂Cl₂ (3×150 mL). The combined extracts were dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica (2% MeOH/CH₂Cl₂) afforded 915 mg (64%) of thedesired aldehyde 36.

Step 2. Methylenation of 36 to Give Alkene 37

The Tebbe reagent (0.5 M in THF, 4.86 mL, 2.43 mmol) was added to asolution of aldehyde 36 (677 mg, 1.65 mmol) in THF (11 mL) at −40° C.under nitrogen. After 1 h at −40° C. the reaction was quenched byaddition of aqueous 2 N NaOH (1.65 mL) and stirred vigorously overnightwith the addition of THF (15 mL). The mixture was filtered throughcelite, washing with excess EtOAc. The filtrate was concentrated invacuo. Purification of the residue by flash column chromatography onsilica (30%→50% EtOAc/Hex) afforded 254 mg (38%) of the desired alkene37.

Step 3. Metathesis Reaction of 37 to Give Alkene 38

Benzylidene[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(tricyclohexylphosphine)-ruthenium(Grubbs' catalyst, 2nd generation, 48 mg, 0.057 mmol) was added to asolution of alkene 37 (230 mg, 0.56 mmol) and methyl5-allylthiophene-2-carboxylate (preparation 3, 206 mg, 1.13 mmol) inCH₂Cl₂ (3.0 mL). The reaction mixture was heated at reflux for 4 h. Thereaction mixture was cooled to rt and more catalyst (48 mg, 0.057 mmol)and methyl 5-allylthiophene-2-carboxylate (100 mg, 0.55 mmol) wereadded. The mixture was heated for 18 h longer at reflux then cooled andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (5%→50% EtOAc/Hex, gradient) afforded 100 mg(32%) of the desired alkene 38 along with 130 mg (57%) of the startingalkene 37.

Step 4. Oxidative Deprotection of 38 to Give 39 and 40

DDQ (58 mg, 0.26 mmol) was added to a mixture of 38 (130 mg, 0.23 mmol)in CH₂Cl₂ (3.1 mL) and water (0.16 mL) at 0° C. under nitrogen. After 45min, the reaction was quenched with saturated aqueous NaHCO₃ (40 mL).The mixture was extracted with EtOAc (3×30 mL). The combined extractswere washed with brine (25 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (50%→75% EtOAc/Hex, gradient) afforded 28 mg ofan inseparable mixture of starting material 38 and ketone 39, and 63 mg(62%) of the desired alcohol 40.

Step 5. Saponification of 40 to Give 41

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofester 40 (3.7 mg, 0.008 mmol) in MeCN (0.2 mL) and pH 7.2 buffer (2.5mL). After 15.5 h at rt, the reaction was diluted with MeCN (8 mL) andconcentrated in vacuo. The residue was suspended in 10% MeOH/CH₂Cl₂ andfiltered through a cotton plug. The filtrate was concentrated in vacuoto afford 3.0 mg (84%) of the title compound (41).

EXAMPLE 175-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (43)

Step 1. Hydrogenation of 40 to Give Ester 42

Palladium on carbon (10 wt. %, 15 mg) was added to a solution of alkene40 (63 mg, 0.14 mmol) in methanol (3.0 mL). A hydrogen atmosphere wasestablished by evacuating and refilling with hydrogen (3×) and thereaction mixture was stirred under a balloon of hydrogen. After 3 h atrt, the reaction mixture was filtered through celite, washing with MeOH,and the filtrate was concentrated in vacuo to afford 63 mg crudeproduct. ¹H NMR analysis showed starting material remaining so the crudematerial was resubmitted to the conditions above. After 20 h at rt, thereaction mixture was filtered through celite, washing with MeOH, and thefiltrate was concentrated in vacuo to afford 60 mg (95%) of the desiredester 42.

Step 2. Saponification of 42 to give 43.

Aqueous 1 N lithium hydroxide (0.19 mL, 0.19 mmol) was added to asolution of ester 42 (17 mg, 0.038 mmol) in THF (0.38 mL). After 20 h atrt, H₂O (1.0 mL) was added and the mixture was acidified with 1 Naqueous HCl (1.0 mL) and extracted with EtOAc (3×10 mL). The combinedextracts were washed with brine (10 mL) then dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica (EtOAc→25% MeOH/EtOAc, gradient) afforded 14.4mg (87%) of the title compound (43).

EXAMPLE 185-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid isopropyl ester (44)

DBU (5.2 μL, 0.035 mmol) was added to a solution of acid 43 (7.5 mg,0.017 mmol) in acetone (0.1 mL) at rt under nitrogen. After 10 min,2-iodopropane (35 μL, 0.35 mmol) was added. After 21 h at rt, thereaction was quenched with 0.01 N HCl (3 mL) and extracted with EtOAc(3×4 mL). The combined extracts were washed with brine (5 mL), dried(Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by flash column chromatography on silica (CH₂Cl₂→1% MeOH/CH₂Cl₂)afforded 4.6 mg (53%) of the title compound (44).

EXAMPLE 195-{3-[(S)-1-(4-Hexanoyl-phenyl)-5-oxo-pyrrolidin-2-yl]-propyl}-thiophene-2-carboxylicacid (46)

Step 1. Oxidation of 38/39 Afford 39

DDQ (5.5 mg, 0.024 mmol) was added to the mixture of ether 38 and ketone39 from Example 16, step 4 (6.8 mg, 0.012 mmol) in CH₂Cl₂ and water(20:1, 0.25 mL) at rt under nitrogen. After 1.5 h, the reaction wasquenched with saturated aqueous NaHCO₃ (5 mL). The mixture was extractedwith EtOAc (3×5 mL). The combined extracts were washed with brine (5 mL)then dried (Na₂SO₄), filtered and concentrated in vacuo. Purification ofthe residue by preparative thin layer chromatography (60% EtOAc/Hex)afforded 1.5 mg (28%) of desired ketone 39.

Step 2. Hydrogenation of 39 to Give Ester 45

Palladium on carbon (10 wt. %, 1 mg) was added to a solution of alkene39 (1.5 mg, 0.0034 mmol) in methanol (0.5 mL). A hydrogen atmosphere wasestablished by evacuating and refilling with hydrogen (3×) and thereaction mixture was stirred under a balloon of hydrogen. After 2 h atrt, the reaction mixture was filtered through celite, washing with MeOH,and the filtrate was concentrated in vacuo to afford 1.3 mg (86%) ofdesired ester 45.

Step 3. Saponification of 45 to Give 46

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofester 45 (1.3 mg, 0.0029 mmol) in MeCN (0.1 mL) and pH 7.2 buffer (2.5mL). After 23 h at rt, the reaction was diluted with MeCN (10 mL) andconcentrated in vacuo. The residue was suspended in 10% MeOH/CH₂Cl₂ andfiltered through a cotton plug. The filtrate was concentrated in vacuoto afford 1.2 mg (95%) of the title compound (46).

EXAMPLE 205-[(R)-1-(4-Hexanoyl-phenyl)-5-oxo-pyrrolidin-2-ylmethoxymethyl]-thiophene-2-carboxylicacid (47)

Rabbit liver esterase (134 units/mg, 1 mg) was added to a solution ofester 27 (Example 10, step 4, 6.6 mg, 0.014 mmol) in MeCN (0.1 mL) andpH 7.2 buffer (2.5 mL). After 17 h at rt, the reaction was diluted withMeCN (8 mL) and concentrated in vacuo. Purification of the residue bypreparative thin layer chromatography (4% MeOH/CH₂Cl₂) afforded 1 mg(17%) of the title compound (47).

EXAMPLES 21 AND 225-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (faster eluting diastereomer 48) and5-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (slower eluting diastereomer 49)

The two diastereomers of example 10 (28, ˜100 mg) were separated on aWaters 600 HPLC instrument employing a Waters 2996 PDA detector and aWhatman Partisil® 10 M20/50 column, 22 mm×500 mm (Cat. No. 4232-220,Q.A. No. 3TA02D80). Using 60% EtOAc/Hex as the eluent and a flow rate of15 mL/min, the first diastereomer (48, 32.8 mg total isolated) eluted at55-60 min, and the second diastereomer (49, 52.6 mg total isolated)eluted at 61-70 min.

EXAMPLE 235-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (50)

Aqueous 1 N lithium hydroxide (0.05 mL, 0.05 mmol) was added to asolution of faster eluting ester diastereomer 48 (2.7 mg, 0.0057 mmol)in THF (0.1 mL) and the mixture was heated at reflux overnight. After 17h, the reaction was cooled to rt, acidified with 0.05 N aqueous HCl (5mL) and extracted with CH₂Cl₂ (3×5 mL). The combined extracts were dried(Na₂SO₄), filtered and concentrated in vacuo to afford 2.5 mg (100%) ofthe title compound (50).

EXAMPLE 245-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (51)

Aqueous 1 N lithium hydroxide (0.05 mL, 0.05 mmol) was added to asolution of slower eluting ester diastereomer 49 (2.8 mg, 0.0059 mmol)in THF (0.1 mL) and the mixture was heated at reflux overnight. After 23h, the reaction was cooled to rt, acidified with 0.05 N aqueous HCl (5mL) and extracted with CH₂Cl₂ (3×5 mL). The combined extracts were dried(Na₂SO₄), filtered and concentrated in vacuo to afford 1.7 mg (67%) ofthe title compound (51).

EXAMPLES 25 AND 265-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid methyl ester (faster eluting diastereomer 52) and5-(3-{(S)-1-[4-(1-hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid methyl ester (slower eluting diastereomer 53)

The two diastereomers from example 17, step 1 (42, ˜43 mg) wereseparated on a Waters 600 HPLC instrument employing a Waters 2996 PDAdetector and a Whatman Partisil® 10 M20/50 column, 22 mm×500 mm (Cat.No. 4232-220, Q.A. No. 3TA02D80). Using 55% EtOAc/Hex as the eluent anda flow rate of 15 mL/min, the first diastereomer (52, 16 mg) eluted at69-75 min, and the second diastereomer (53, 19 mg) eluted at 80-88 min.

EXAMPLE 275-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (54)

Rabbit liver esterase (134 units/mg, 2 mg) was added to a solution offaster eluting ester diastereomer 52 (16 mg, 0.036 mmol) in MeCN (0.2mL) and pH 7.2 buffer (3.0 mL). After 18 h at rt, the reaction wasdiluted with MeCN (10 mL) and concentrated in vacuo. The residue wasdiluted with CH₂Cl₂ (5 mL), filtered through a plug of glass wool andconcentrated in vacuo. Purification of the residue by preparative thinlayer chromatography (EtOAc→25% MeOH/EtOAc, gradient) afforded 12 mg(77%) of the title compound (54).

EXAMPLE 285-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (55)

Rabbit liver esterase (134 units/mg, 2 mg) was added to a solution ofslower eluting ester diastereomer 53 (19 mg, 0.043 mmol) in MeCN (0.2mL) and pH 7.2 buffer (3.0 mL). After 18 h at rt, the reaction wasdiluted with MeCN (10 mL) and concentrated in vacuo. The residue wasdiluted with CH₂Cl₂ (5 mL), filtered through a plug of glass wool andconcentrated in vacuo. Purification of the residue by preparative thinlayer chromatography (EtOAc→25% MeOH/EtOAc, gradient) afforded 10.5 mg(57%) of the title compound (55).

EXAMPLE 295-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid isopropyl ester (56, from 54 and 52)

DBU (4.2 μL, 0.028 mmol) and 2-iodopropane (19 μL, 0.19 mmol) were addedto a solution of acid 54 (8 mg, 0.019 mmol) in acetone (0.15 mL) at rtunder nitrogen. After 18 h at rt, the solvent was removed under a streamof nitrogen. The residue was diluted with EtOAc (10 mL) and washed with0.1 N HCl (2×5 mL) and brine (5 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (CH₂Cl₂→5% MeOH/CH₂Cl₂) afforded 1.9 mg (22%)of the title compound (56) and 4 mg (50%) recovered 54.

EXAMPLE 305-(3-{(S)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid isopropyl ester (57, from 55 and 53)

DBU (4.7 μL, 0.031 mmol) and 2-iodopropane (21 μL, 0.21 mmol) were addedto a solution of acid 55 (9 mg, 0.021 mmol) in acetone (0.2 mL) at rtunder nitrogen. After 18 h at rt, the solvent was removed under a streamof nitrogen. The residue was diluted with EtOAc (10 mL) and washed with0.1 N HCl (2×5 mL) and brine (5 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (CH₂Cl₂→25% MeOH/CH₂Cl₂) afforded 2.0 mg (20%)of the title compound (57) and 6 mg (67%) recovered 55.

EXAMPLE 315-{(R)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (61)

Step 1. Arylation of 1 to Give 58

Pd₂(dba)₃ (550 mg, 0.60 mmol), xantphos (1.04 g, 180 mmol) and Cs₂CO₃(5.87 g, 18.0 mmol) were added sequentially to a solution of amide 1(3.45 g, 15.0 mmol) in 1,4-dioxane (100 mL). A solution of1-(1-(4-methoxybenzyloxyheptyl)-4-bromobenzene (preparation 4, 5.30 g,13.54 mmol) in 1,4-dioxane (50 mL) was added via cannula. The reactionmixture was purged with nitrogen then heated at reflux overnight. After17 h, the reaction was cooled to rt and filtered through celite, washingwith CH₂Cl₂. The filtrate was concentrated in vacuo and the residue waspurified by flash column chromatography on silica gel (5%→35%EtOAc/Hexane, gradient) to afford 5.26 g (72%) of the desired product58.

Step 2. Deprotection of 58 to Give 59

HF-pyridine (8.8 mL) was added to a solution of silyl ether 58 (5.26 g,9.74 mmol) in MeCN (50 mL) in a plastic bottle at 0° C. After 45 min at0° C., the reaction was quenched with saturated aqueous NaHCO₃ (400 mL).The mixture was extracted with EtOAc (3×200 mL). The combined extractswere washed with brine (200 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (CH₂Cl₂→5% MeOH/CH₂Cl₂, gradient) afforded 3.9g (94%) of the desired alcohol 59 as a pale yellow solid.

Step 3. Alkylation of 59 to Give 60

A round bottom flask was charged with potassium hydride (30 wt % in oil,138 mg, 1.03 mmol). The material was washed with hexanes (3×1 mL), thensuspended in THF (1 mL). The mixture was cooled to 0° C. and a solutionof alcohol 59 (339 mg, 0.80 mmol) in THF (1.5 mL) was added via cannula.After 1 h at 0° C., a solution of isopropyl5-chloromethylthiophene-2-carboxylate (preparation 2, 174 mg, 0.80 mmol)in THF (1.5 mL) was added via cannula. Potassium iodide (14 mg, 0.08mmol) was added and the reaction was allowed to warm to rt. After 18 h,the reaction was quenched with saturated aqueous NH₄Cl (15 mL) andextracted with EtOAc (3×25 mL). Combined extracts were washed with brine(15 mL) then dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(20%→75% EtOAc/Hexane, gradient), followed by preparative thin layerchromatography (65% EtOAc/Hexane) afforded 65 mg (14%) of desiredproduct 60.

Step 4. Oxidative Deprotection of 60 to Give 61

DDQ (26 mg, 0.12 mmol) was added to a solution of 60 (65 mg, 0.11 mmol)in CH₂Cl₂ (1.4 mL) and water (0.07 mL) at 0° C. under nitrogen. After 40min, the reaction was quenched with saturated aqueous NaHCO₃ (20 mL) andextracted with EtOAc (3×20 mL). Combined extracts were washed with brine(15 mL) then dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(50%→75% EtOAc/Hexane, gradient), followed by preparative thin layerchromatography (60% EtOAc/Hexane) afforded 36 mg (69%) of the titlecompound (61).

EXAMPLES 32 AND 335-{(R)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (faster eluting diastereomer 62) and5-{(R)-1-[4-(1-hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid isopropyl ester (slower eluting diastereomer 63)

The two diastereomers of example 31 (61, ˜36 mg) were separated on aWaters 600 HPLC instrument employing a Waters 2996 PDA detector and aWhatman Partisil® 10 M20/50 column, 22 mm×500 mm (Cat. No. 4232-220,Q.A. No. 3TA02D80). Using 60% EtOAc/Hex as the eluent and a flow rate of15 mL/min, the first diastereomer (62, 14.8 mg) eluted at 50-56.5 min,and the second diastereomer (63, 16.4 mg) eluted at 56.5-70 min.

EXAMPLE 345-{(R)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (64)

Aqueous 1 N lithium hydroxide (0.05 mL, 0.05 mmol) was added to asolution of faster eluting ester diastereomer 62 (3.5 mg, 0.0072 mmol)in THF (0.1 mL) and the mixture was heated at reflux overnight. After 18h, the reaction was cooled to rt, diluted with water (2 mL), acidifiedwith 1.0 N aqueous HCl (1 mL) and extracted with EtOAc (3×5 mL). Thecombined extracts were washed with brine (5 mL), dried (Na₂SO₄),filtered and concentrated in vacuo to afford 3.0 mg (94%) of the titlecompound (64).

EXAMPLE 355-{(R)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxymethyl}-thiophene-2-carboxylicacid (65)

Aqueous 1 N lithium hydroxide (0.05 mL, 0.05 mmol) was added to asolution of slower eluting ester diastereomer 63 (3.5 mg, 0.0072 mmol)in THF (0.1 mL) and the mixture was heated at reflux overnight. After 18h, the reaction was cooled to rt, diluted with water (2 mL), acidifiedwith 1.0 N aqueous HCl (1 mL) and extracted with EtOAc (3×5 mL). Thecombined extracts were washed with brine (5 mL), dried (Na₂SO₄),filtered and concentrated in vacuo to afford 3.2 mg (99%) of the titlecompound (65).

EXAMPLE 365-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (71)

Step 1. Oxidation of 59 to Give Aldehyde 66

1-(3-(Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 1.43g, 7.45 mmol) and DMSO (0.70 mL, 9.86 mmol) were added sequentially to asolution of alcohol 59 (1.06 g, 2.48 mmol) in benzene (25 mL) at rtunder nitrogen. After 10 min at rt, pyridinium trifluoroacetate (527 mg,2.73 mmol) was added. After 3 h at rt, the solution was decanted fromthe oily residue and the residue was washed with benzene (3×15 mL). Thecombined benzene phases were concentrated in vacuo. Purification of theresidue by flash column chromatography on silica (CH₂Cl₂→3% MeOH/CH₂Cl₂,gradient) afforded 1.0 g (95%) of the desired aldehyde 66.

Step 2. Methylenation of 66 to Give Alkene 67

The Tebbe reagent (0.5 M in THF, 7.0 mL, 3.5 mmol) was added to asolution of aldehyde 66 (1.0 g, 2.36 mmol) in THF (16 mL) at −40° C.under nitrogen. After 1 h at −40° C. the reaction was quenched byaddition of aqueous 2 N NaOH (5.25 mL) and stirred vigorously overnightwith the addition of THF (20 mL). The mixture was filtered throughcelite, washing with excess EtOAc. The filtrate was concentrated invacuo. Purification of the residue by flash column chromatography onsilica (40% EtOAc/Hex) afforded 195 mg (20%) of the desired alkene 67.

Step 3. Metathesis Reaction of 67 to Give Alkene 68

Grubbs' second generation catalyst (38 mg, 0.045 mmol) was added to asolution of alkene 67 (190 mg, 0.45 mmol) and methyl5-allylthiophene-2-carboxylate (preparation 3, 173 mg, 0.95 mmol) inCH₂Cl₂ (2.4 mL). The reaction mixture was heated at reflux for 2 h. Thereaction mixture was cooled to rt and more catalyst (9 mg, 0.011 mmol)and methyl 5-allylthiophene-2-carboxylate (165 mg, 0.91 mmol) wereadded. The mixture was heated for 22 h longer at reflux then cooled andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (2 times, first using 5%→50% EtOAc/Hex,gradient then second using CH₂Cl₂→3% MeOH/CH₂Cl₂, gradient) afforded 180mg (69%) of the desired alkene 68.

Step 4. Oxidative Deprotection of 68 to Give 69

DDQ (78 mg, 0.34 mmol) was added to a mixture of 68 (180 mg, 0.31 mmol)in CH₂Cl₂ (4.1 mL) and water (0.21 mL) at 0° C. under nitrogen. After 45min at 0° C., the reaction was quenched with saturated aqueous NaHCO₃(50 mL). The mixture was extracted with EtOAc (3×50 mL). The combinedextracts were washed with brine (50 mL) then dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica (50%→66% EtOAc/Hex, gradient) afforded 50 mg(35%) of the desired alcohol 69.

Step 5. Hydrogenation of 69 to Give Ester 70

Palladium on carbon (10 wt. %, 12 mg) was added to a solution of alkene69 (50 mg, 0.11 mmol) in methanol (2.3 mL). A hydrogen atmosphere wasestablished by evacuating and refilling with hydrogen (3×) and thereaction mixture was stirred under a balloon of hydrogen. After 20 h atrt, the reaction mixture was filtered through celite, washing with MeOH,and the filtrate was concentrated in vacuo to afford 50 mg (99%) of thedesired ester 70.

Step 6. Saponification of 70 to Give 71

Aqueous 1 N lithium hydroxide (0.19 mL, 0.19 mmol) was added to asolution of ester 70 (17 mg, 0.038 mmol) in THF (0.4 mL). After 18 h atrt, H₂O (1.0 mL) was added and the mixture was acidified with 1 Naqueous HCl (1.0 mL) and extracted with EtOAc (3×10 mL). The combinedextracts were washed with brine (10 mL) then dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by preparativethin layer chromatography (15% MeOH/CH₂Cl₂) afforded 5.6 mg (34%) of thetitle compound (71).

EXAMPLES 37 AND 385-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid methyl ester (faster eluting diastereomer 72) and5-(3-{(S)-1-[4-(1-hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid methyl ester (slower eluting diastereomer 73)

The two diastereomers from example 36, step 5 (70, ˜34 mg) wereseparated on a Waters 600 HPLC instrument employing a Waters 2996 PDAdetector and a Whatman Partisil® 10 M20/50 column, 22 mm×500 mm (Cat.No. 4232-220, Q.A. No. 3TA02D80). Using 55% EtOAc/Hex as the eluent anda flow rate of 15 mL/min, the first diastereomer (72, 10.7 mg) eluted at78-87.5 min, and the second diastereomer (73, 7.0 mg) eluted at 91-101min.

EXAMPLE 395-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (74)

Aqueous 1 N lithium hydroxide (0.12 mL, 0.12 mmol) was added to asolution of faster eluting ester diastereomer 72 (10.7 mg, 0.023 mmol)in THF (0.3 mL). After 66 h at rt, H₂O (1.0 mL) was added and themixture was acidified with 1 N aqueous HCl (1.0 mL) and extracted withEtOAc (3×10 mL). The combined extracts were washed with brine (5 mL)then dried (Na₂SO₄), filtered and concentrated in vacuo to afford 10 mg(96%) of the title compound (74).

EXAMPLE 405-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid (75)

Aqueous 1 N lithium hydroxide (0.08 mL, 0.08 mmol) was added to asolution of slower eluting ester diastereomer 73 (7.0 mg, 0.015 mmol) inTHF (0.2 mL). After 66 h at rt, H₂O (1.0 mL) was added and the mixturewas acidified with 1 N aqueous HCl (1.0 mL) and extracted with EtOAc(3×8 mL). The combined extracts were washed with brine (5 mL) then dried(Na₂SO₄), filtered and concentrated in vacuo to afford 6.5 mg (96%) ofthe title compound (75).

EXAMPLE 415-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid isopropyl ester (76, from 74 and 72)

DBU (4.0 μL, 0.027 mmol) and 2-iodopropane (36 μL, 0.36 mmol) were addedto a solution of acid 74 (8 mg, 0.018 mmol) in acetone (0.2 mL) at rtunder nitrogen. After 72 h at rt, the solvent was removed under a streamof nitrogen. The residue was diluted with EtOAc (10 mL) and washed with0.5 N HCl (2×5 mL) and brine (5 mL) then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica (EtOAc→20% MeOH/EtOAc) afforded 7.3 mg (83%) ofthe title compound (76).

EXAMPLE 425-(3-{(S)-1-[4-(1-Hydroxy-heptyl)-phenyl]-5-oxo-pyrrolidin-2-yl}-propyl)-thiophene-2-carboxylicacid isopropyl ester (77, from 75 and 73)

DBU (2.5 μL, 0.017 mmol) and 2-iodopropane (22.5 μL, 0.225 mmol) wereadded to a solution of acid 75 (5 mg, 0.011 mmol) in acetone (0.11 mL)at rt under nitrogen. After 72 h at rt, the solvent was removed under astream of nitrogen. The residue was diluted with EtOAc (10 mL) andwashed with 0.5 N HCl (2×5 mL) and brine (5 mL) then dried (Na₂SO₄),filtered and concentrated in vacuo. Purification of the residue by flashcolumn chromatography on silica (EtOAc→20% MeOH/EtOAc) afforded 3.2 mg(58%) of the title compound (77).

EXAMPLE 434-{(R)-1-[4-(1-Hydroxy-hexyl)-phenyl]-5-oxo-pyrrolidin-2-ylmethoxy}-benzoicacid (80)

Step 1. Mitsunobu Reaction of 25 and Methyl 4-Hydroxybenzoate to Give 78

Diisopropyl azodicarboxylate (DIAD, 194 μL, 1.0 mmol) was added to asolution of alcohol 25 (200 mg, 0.49 mmol), triphenylphosphine (191 mg,0.73 mmol) and methyl 4-hydroxybenzoate (87 mg, 0.57 mmol) in CH₂Cl₂(2.5 mL). After stirring 18 h at rt, the solvent was removed under astream of nitrogen and the residue was suspended in EtOAc (75 mL). Themixture was washed with saturated aqueous NaHCO₃ (3×25 mL) and brine (25mL) then the organic phase was dried (Na₂SO₄) filtered and concentratedin vacuo. Purification of the residue by flash column chromatography onsilica gel (50% EtOAc/hexane →EtOAc, gradient) afforded 81 mg (31%) ofthe desired ether 78.

Step 2. Oxidative Deprotection of 78 to Give 79

DDQ (37 mg, 0.16 mmol) was added to a mixture of 78 (81 mg, 0.15 mmol)in CH₂Cl₂ (2.0 mL) and water (0.1 mL) at 0° C. under nitrogen. After 45min at 0° C., the reaction was quenched with saturated aqueous NaHCO₃(25 mL). The mixture was extracted with EtOAc (3×25 mL). The combinedextracts were washed with brine (25 mL) then dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica (85% EtOAc/Hex→EtOAc, gradient) afforded 31 mg(49%) of the desired alcohol 79.

Step 3. Saponification of 79 to Give 80

Aqueous 1 N lithium hydroxide (0.35 mL, 0.35 mmol) was added to asolution of ester 79 (30 mg, 0.071 mmol) in THF (0.7 mL). After 20 h atrt, water (2.0 mL) was added and the mixture was acidified with 1 Naqueous HCl (1.5 mL) and extracted with EtOAc (3×10 mL). The combinedextracts were washed with brine (10 mL) then dried (Na₂SO₄), filteredand concentrated in vacuo. Purification of the residue by flash columnchromatography on silica (EtOAc→10% MeOH/EtOAc, gradient) afforded 11.5mg (38%) of starting ester 79 and 8.5 mg (29%) of the title compound(80).

Preparation 1 1-(1-(4-Methoxybenzyloxy-hexyl)-4-bromobenzene

Step 1. Pentyl Grignard Addition to 4-Bromobenzaldehyde

n-Pentyl magnesium bromide (2.0 M in THF, 27 mL, 54 mmol) was added to asolution of 4-bromobenzaldehyde (5.0 g, 27 mmol) in THF (20 mL) at 0° C.under nitrogen. After 1 h, the reaction was quenched with 3 N HCl andextracted with Et₂O (3×120 mL). Combined extracts were washed with brine(100 mL), dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica gel(5% EtOAc/Hex) afforded 5.1 g (74%) of 1-(4-bromophenyl)-hexan-1-ol.

Step 2. Protection of the Alcohol as its Mpm Ether

Sodium hydride (60% wt. in oil, 0.95 g, 23.8 mmol) was added to asolution of the alcohol from step 1 (5.11 g, 19.9 mmol) in THF and DMF(2:1, 20 mL) at 0° C. under nitrogen. After 1 h at 0° C.,4-methoxybenzyl chloride (3.23 mL, 23.8 mmol) and the reaction wasallowed to warm to rt. The reaction was then heated at 80° C. After 17h, the reaction was allowed to cool to rt, quenched with saturatedaqueous NH₄Cl (100 mL) and extracted with EtOAc (3×100 mL). The combinedextracts were washed with brine (100 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (2% EtOAc/Hex) afforded 7.02 g (94%) of thetitle compound.

Preparation 2 Isopropyl 5-chloromethylthiophene-2-carboxylate

Step 1. Preparation of the Bis-Isopropyl Ester

DBU (31.3 mL, 209 mmol) and 2-iodopropane (20.9 mL, 209 mmol) were addedto a solution of thiophene-2,5-dicarboxylic acid (6.0 g, 34.9 mmol) inacetone (60 mL) at rt under nitrogen. After 21 h at rt, the reaction wasquenched with saturated aqueous NaHCO₃ (300 mL) and extracted with EtOAc(3×150 mL). The combined extracts were washed with brine (200 mL), dried(Na₂SO₄), filtered and concentrated in vacuo to afford 7.59 g (85%) ofthe diester.

Step 2. Reduction to the Hydroxymethyl Ester

Sodium borohydride (3.36 g, 88.8 mmol) was added to a solution of thediester (7.59 g, 29.6 mmol) in CH₂Cl₂/MeOH (1:1, 100 mL) at 0° C. undernitrogen. The ice bath was removed and the reaction was allowed to stirat rt overnight. After 20.5 h at rt the reaction was concentrated invacuo then aqueous 0.5 N HCl (100 mL) was added. The mixture wasextracted with CH₂Cl₂ (3×100 mL). The combined extracts were dried(Na₂SO₄), filtered and concentrated in vacuo. Purification of theresidue by flash column chromatography on silica (5%→60% EtOAc/Hex,gradient) afforded 738 mg (12%) of the alcohol.

Step 3. Conversion of the Alcohol to the Chloride

Methanesulfonyl chloride (0.67 mL, 8.1 mmol) and triethylamine (1.7 mL,12.2 mmol) were added sequentially and dropwise to a solution of thealcohol (696 mg, 3.48 mmol) in CH₂Cl₂ (4.0 mL) at 0° C. under nitrogen.The ice bath was removed and the reaction was allowed to stir overnightat rt. After 17 h, the reaction was quenched with saturated aqueousNaHCO₃ (30 mL) and extracted with CH₂Cl₂ (3×50 mL). The combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo.Purification of the residue by flash column chromatography on silica (5%EtOAc/Hex) afforded 664 mg (87%) of the title compound.

Preparation 3 Methyl 5-allylthiophene-2-carboxylate

Step 1. Preparation of the Methyl Ester

Acetyl chloride (6.9 mL, 96.6 mmol) was added to a solution of5-bromo-2-thiophenecarboxylic acid (4.0 g, 19.3 mmol) in methanol (30mL) at rt. After 17 h at rt, the reaction was heated at reflux for 1.5 hto drive it to completion. The reaction was then cooled to rt andconcentrated in vacuo to remove methanol. Saturated aqueous NH₄Cl (120mL) was added and the mixture was extracted with CH₂Cl₂ (3×100 mL). Thecombined extracts were dried (Na₂SO₄), filtered and concentrated invacuo to afford 3.57 g (84%) of the desired methyl ester as an off whitesolid.

Step 2. Allylation of the Bromothiophene

Isopropyl magnesium chloride (2.0 M in Et₂O, 8.9 mL, 17.8 mmol) wasadded to a solution of the bromide from step 1 (3.56 g, 16.1 mmol) inTHF (10 mL) at −40° C. under nitrogen. The reaction mixture was stirredat −40° C. for 1 h, then copper (I) cyanide (144 mg, 1.61 mmol) andallyl bromide (3.0 mL, 35.4 mmol) were added sequentially. The reactionmixture was stirred at −40° C. for 1 h then was quenched with saturatedaqueous NH₄Cl (100 mL) and extracted with EtOAc (3×100 mL). The combinedextracts were washed with brine (100 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (5% EtOAc/Hex) afforded 2.45 g (83%) of thetitle compound as a pale yellow oil that solidified on standing.

Preparation 4 1-(1-(4-Methoxybenzyloxy-heptyl)-4-bromobenzene

Step 1. Hexyl Grignard Addition to 4-Bromobenzaldehyde

n-Hexyl magnesium bromide (2.0 M in Et₂O, 27 mL, 54 mmol) was added to asolution of 4-bromobenzaldehyde (5.0 g, 27 mmol) in THF (20 mL) at 0° C.under nitrogen. After 1.5 h at 0° C., the reaction was quenched slowlywith 3 N HCl (20 mL) and concentrated in vacuo. The residue was dilutedwith water (30 mL) and extracted with Et₂O (3×150 mL). Combined extractswere dried (Na₂SO₄), filtered and concentrated in vacuo. Purification ofthe residue by flash column chromatography on silica gel (5%→10%EtOAc/Hex) afforded 5.6 g (76%) of 1-(4-bromophenyl)-heptan-1-ol.

Step 2. Protection of the Alcohol as its MPM Ether

Sodium hydride (60% wt. in oil, 0.991 g, 24.8 mmol) was added to asolution of the alcohol from step 1 (5.6 g, 20.6 mmol) in THF and DMF(2:1, 30 mL) at 0° C. under nitrogen. After 5 min at 0° C., the reactionwas allowed to warm to rt and 4-methoxybenzyl chloride (3.4 mL, 25.0mmol) was added. The reaction was then heated at 80° C. After 18 h at80° C., the reaction was allowed to cool to rt, quenched with saturatedaqueous NH₄Cl (50 mL) and concentrated in vacuo. The remainder wasextracted with EtOAc (3×100 mL). The combined extracts were washed withwater (2×100 mL) and brine (75 mL), then dried (Na₂SO₄), filtered andconcentrated in vacuo. Purification of the residue by flash columnchromatography on silica gel (2% EtOAc/Hex) afforded 7.5 g (93%) of thetitle compound.

EP2 data EP4 data flipr cAMP Ki pH flipr Other Receptors (EC50 in nM)Example# Structure EC50 EC50 Ki 6.0 EC50 KI hFP hEP1 hEP3A hTP hIP hDP11

>10000 517 NA NA >10000 NA NA NA NA NA NA 12

212 8 387 NA >10000 NA NA NA NA NA NA 13

20 1.5 190 NA >10000 NA NA 519 NA NA 5763 14

426 27 1639 NA >10000 NA NA NA NA NA NA 15

1812 312 5731 >10000 >10000 NA NA NA NA NA 7560 16

226 15 1382 NA >10000 NA NA 1411 NA NA NA 17

5 0.55 23 NA >10000 NA NA  155 NA NA 1234 19

16 1.6 31 >10000 >10000 NA NA 2345 NA NA 7695 20

215 8 163 NA >10000 NA NA NA NA NA NA 23

62 5 345 >10000 >10000 NA NA  153 NA NA 7749 24

15 1.5 116 >10000    6032 NA NA 1205 NA NA 6800 27

6 0.19 21 >10000 >10000 NA NA  12 NA NA  812 28

1.6 0.15 15 >10000   4849 NA NA  156 NA NA  296 34

134 7 229 NA   3842 NA NA  71 NA NA 6829 35

49 4 201 NA 3288 NA NA  621 NA NA NA 36

30 0.9 10 >10000 NA NA  47 NA NA  105 39

16 1.4 12 NA NA 6952   7 NA NA  37 40

5 0.6 8 NA NA NA  33 NA >10000  106 43

7284 NA 22726 NA 8752 NA NA NA NA NA NA

Biological Assay Methods

Binding Data

Ki

Competition binding experiments were performed in a medium containingHank's balanced salt solution, Hepes 20 mM, pH 7.3, membranes (˜60 μgprotein) or 2×10⁵ cells from HEK 293 cells stably expressing human EP2receptors, [³H]PGE2 (10 nM) and various concentrations of test compoundsin a total volume of 300 μl. Reaction mixtures were incubated at 23° C.for 60 min, and were filtered over Whatman GF/B filters under vacuum.Filters were washed three times with 5 ml ice-cold buffer containing 50mM Tris/HCl (pH 7.3). Non-specific binding was estimated in the presenceof excess unlabeled PGE2 (10 μM). Binding data fitted to the bindingmodel for a single class of binding sites, using nonlinear regressionanalysis. IC₅₀ values thus obtained were converted to Ki using theequation of Ki=(IC₅₀/(1+[L]/K_(D)) where [L] represents PGE2concentration (10 nM) and K_(D) the dissociation constant for [³H]PGE2at human EP2 receptors (40 nM).

Radioligand Binding

Cells Stably Expressing EP₁, EP₂, EP₄ and FP Receptors

HEK-293 cells stably expressing the human or feline FP receptor, or EP₁,EP₂, or EP₄ receptors were washed with TME buffer, scraped from thebottom of the flasks, and homogenized for 30 sec using a Brinkman PT10/35 polytron. TME buffer was added to achieve a final 40 ml volume inthe centrifuge tubes (the composition of TME is 100 mM TRIS base, 20 mMMgCl₂, 2M EDTA; 10N HCl is added to achieve a pH of 7.4).

The cell homogenate was centrifuged at 19000 r.p.m. for 20 min at 4° C.using a Beckman Ti-60 rotor. The resultant pellet was resuspended in TMEbuffer to give a final 1 mg/ml protein concentration, as determined byBiorad assay. Radioligand binding competition assays vs. [³H-]17-phenylPGF_(2α) (5 nM) were performed in a 1001 volume for 60 min. Bindingreactions were started by adding plasma membrane fraction. The reactionwas terminated by the addition of 4 ml ice-cold TRIS-HCl buffer andrapid filtration through glass fiber GF/B filters using a Brandel cellharvester. The filters were washed 3 times with ice-cold buffer and ovendried for one hour.

[³H-] PGE₂ (specific activity 180 Ci mmol) was used as the radioligandfor EP receptors. [³H] 17-phenyl PGF₂, was employed for FP receptorbinding studies. Binding studies employing EP₁, EP₂, EP₄ and FPreceptors were performed in duplicate in at least three separateexperiments. A 200 μl assay volume was used. Incubations were for 60 minat 25° C. and were terminated by the addition of 4 ml of ice-cold 50 mMTRIS-HCl, followed by rapid filtration through Whatman GF/B filters andthree additional 4 ml washes in a cell harvester (Brandel). Competitionstudies were performed using a final concentration of 5 nM [³H]-PGE₂, or5 nM [³H] 17-phenyl PGF₂, and non-specific binding determined with 10⁻⁵Mof unlabeled PGE₂, or 17-phenyl PGF_(2α), according to receptor subtypestudied.

Methods for FLIPR™ Studies

(a) Cell Culture

HEK-293(EBNA) cells, stably expressing one type or subtype ofrecombinant human prostaglandin receptors (prostaglandin receptorsexpressed: hDP/Gqs5; hEP₁; hEP₂/Gqs5; hEP_(3A)/Gqi5; hEP₄/Gqs5; hFP;hIP; hTP), were cultured in 100 mm culture dishes in high-glucose DMEMmedium containing 10% fetal bovine serum, 2 mM 1-glutamine, 250 μg/mlgeneticin (G418) and 200 μg/ml hygromycin B as selection markers, and100 units/ml penicillin G, 100 μg/ml streptomycin and 0.25 μg/mlamphotericin B.

(b) Calcium Signal Studies on The FLIPR™

Cells were seeded at a density of 5×10⁴ cells per well in Biocoat®Poly-D-lysine-coated black-wall, clear-bottom 96-well plates(Becton-Dickinson) and allowed to attach overnight in an incubator at37° C. Cells were then washed two times with HBSS-HEPES buffer (HanksBalanced Salt Solution without bicarbonate and phenol red, 20 mM HEPES,pH 7.4) using a Denley Cellwash plate washer (Labsystems). After 45minutes of dye-loading in the dark, using the calcium-sensitive dyeFluo-4 μM at a final concentration of 2 μM, plates were washed fourtimes with HBSS-HEPES buffer to remove excess dye leaving 100 μl in eachwell. Plates were re-equilibrated to 37° C. for a few minutes.

Cells were excited with an Argon laser at 488 nm, and emission wasmeasured through a 510-570 nm bandwidth emission filter (FLIPR™,Molecular Devices, Sunnyvale, Calif.). Drug solution was added in a 50μl volume to each well to give the desired final concentration. The peakincrease in fluorescence intensity was recorded for each well. On eachplate, four wells each served as negative (HBSS-HEPES buffer) andpositive controls (standard agonists: BW245C (hDP); PGE₂ (hEP₁;hEP₂/Gqs5; hEP_(3A)/Gqi5; hEP₄/Gqs5); PGF_(2α) (hFP); carbacyclin (hIP);U-46619 (hTP), depending on receptor). The peak fluorescence change ineach drug-containing well was then expressed relative to the controls.

Compounds were tested in a high-throughput (HTS) orconcentration-response (CoRe) format. In the HTS format, forty-fourcompounds per plate were examined in duplicates at a concentration of10⁻⁵ M. To generate concentration-response curves, four compounds perplate were tested in duplicates in a concentration range between 10⁻⁵and 10⁻¹¹ M. The duplicate values were averaged. In either, HTS or CoReformat each compound was tested on at least 3 separate plates usingcells from different passages to give an n>3.

Intraocular Pressure (IOP)

Intraocular pressure studies in dogs involve pneumatonometry performedon conscious Beagle dogs of both sexes (10-15 kg). The animals remainconscious throughout the study and are gently restrained by hand. Drugsare administered topically to one eye as a 25 μL volume drop, the othereye receives 25 μL vehicle (0.1% polysorbate 80:10 mM TRIS) as acontrol. Proparacaine (0.1%) is used for corneal anesthesia duringtonometry. Intraocular pressure is determined just before drugadministration and at 2, 4 and 6 hr thereafter on each day of the 5 daystudy. Drug is administered immediately after the first IOP reading.

The results of the binding and activity studies, presented in Table 1below, demonstrate that the compounds disclosed herein are selectiveprostaglandin EP₂ agonists, and are thus useful for the treatment ofglaucoma, ocular hypertension, inflammatory bowel disease, and the otherdiseases or conditions disclosed herein.

The foregoing description details specific methods and compositions thatcan be employed to practice the present invention, and represents thebest mode contemplated. However, it is apparent for one of ordinaryskill in the art that further compounds with the desired pharmacologicalproperties can be prepared in an analogous manner, and that thedisclosed compounds can also be obtained from different startingcompounds via different chemical reactions. Similarly, differentpharmaceutical compositions may be prepared and used with substantiallythe same result. Thus, however detailed the foregoing may appear intext, it should not be construed as limiting the overall scope hereof,rather, the ambit of the present invention is to be governed only by thelawful construction of the appended claims.

1. A compound according to a formula selected from


2. The compound of claim 1 according to the formula

or a pharmaceutically acceptable salt thereof.
 3. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 4. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 5. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 6. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 7. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 9. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 10. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 11. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 12. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 13. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 14. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 15. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 16. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 17. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 18. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.
 19. compound of claim 1according to the formula

or a pharmaceutically acceptable salt thereof.
 20. The compound of claim1 according to the formula

or a pharmaceutically acceptable salt thereof.